小干扰RNA SOX5对胃癌细胞增殖和凋亡的影响

Effect of small interfering RNAsex determining region Y box protein 5 on proliferation and apoptosis of gastric cancer cells

[目的]探讨Y染色体上的性别决定区相关高迁移率族盒蛋白-5(SOX5)的小干扰RNA(siRNA)对胃癌细胞BGC823增殖及凋亡的影响。[方法]通过qRT-PCR和Western-blot分别检测胃癌细胞系(SGC7901,BGC823,AGS,HGC27)及正常胃上皮细胞GES-1中SOX5 mRNA和蛋白的表达情况;取对数生长周期的胃癌细胞BGC823,通过Lipofiectamine2000将设计合成的SOX5的特异性siRNA以及NC转染BGC823细胞,作为siSOX5组和NC组,不做任何处理的细胞作为空白对照组。qRT-PCR检测各组细胞中SOX5 mRNA的表达情况;Western-blot检测各组SOX5及CyclinD1、P21、Bax、Bcl-2蛋白的表达;MTT检测转染12 h、24 h、48 h、72 h后,各组细胞的增殖情况;流式细胞仪检测转染72 h后各组细胞的凋亡情况。[结果]与正常胃上皮细胞GES-1相比,胃癌细胞SGC7901,BGC823,AGS,HGC27中SOX5 mRNA和蛋白表达升高(P<0.05);转染siSOX5后,siSOX5组SOX5 mRNA及蛋白表达降低(P<0.05);MTT结果显示:与空白对照组和NC组相比,siSOX5组细胞增殖受到抑制,且CyclinD1蛋白表达水平显著下降,p21蛋白表达显著升高(P<0.05);流式细胞检测结果显示:与空白对照组和NC组相比,siSOX5组细胞凋亡率升高,且Bax蛋白表达升高,Bcl-2蛋白表达下降(P<0.05)。[结论]干扰SOX5能够降低胃癌BGC823细胞中SOX5的表达,抑制BGC823细胞的增殖并促进凋亡。

[Objective]To investigate the effect of small interfering RNA of sex determining region Y box protein 5(SOX5)on the proliferation and apoptosis of gastric cancer cell BGC823.[Methods]The expression of SOX5 mRNA and protein in gastric cancer cell lines(SGC7901,BGC823,AGS,HGC27)and normal gastric epithelial cells GES-1 were detected by qRT-PCR and Western-blot.BGC823 cells with logarithmic growth cycle were transfected with SOX5 specific siRNA and NC by Lipofiectamine 2000 as siSOX5 group and NC group,and the cells without any treatment were used as blank control group.The expression of SOX5 mRNA was detected by qRT-PCR in three groups of cells.The expression of SOX5 and CyclinD1,P21,Bax,Bcl-2 protein was detected by Western-blot.MTT assay was used to detect the proliferation of the three groups at 12 h,24 h,48 h and 72 h after transfection.Flow cytometry was used to detect the apoptosis of the three groups of cells 72 h after transfection.[Results]Compared with normal gastric epithelial cells GES-1,the expression levels of SOX5 mRNA and protein in gastric cancer cells SGC7901,BGC823,AGS,and HGC27 were increased(P<0.05),while the expression levels of SOX5 mRNA and protein in siSOX5 group were decreased(P<0.05).MTT results showed that compared with the blank control group and NC group,cell proliferation was inhibited,the expression of CyclinD1 protein was significantly decreased,and the expression of P21 protein was significantly increased in siSOX5 group(P<0.05).The results of flow cytometry showed that compared with the blank control group and NC group,the apoptosis rate,the expression of Bax protein were increased,while the expression of Bcl-2 protein was decreased(P<0.05).[Conclusion]Interference with SOX5 can reduce the expression of SOX5 in BGC823 cells,inhibit the proliferation of BGC823 cells and promote apoptosis.